title: Novel real-time polymerase chain reaction assay for simultaneous detection of recurrent fusion genes in acute myeloid leukemia.
abstract: The recent World Health Organization classification recognizes different subtypes of acute myeloid leukemia (AML) according to the presence of several recurrent genetic abnormalities. Detection of these abnormalities and other molecular changes is of increasing interest because it contributes to a refined diagnosis and prognostic assessment in AML and enables monitoring of minimal residual disease. These genetic abnormalities can be detected using single RT-PCR, although the screening is still labor intensive and costly. We have developed a novel real-time RT-PCR assay to simultaneously detect 15 AML-associated rearrangements that is a simple and easily applicable method for use in clinical diagnostic laboratories. This method showed 100% specificity and sensitivity (95% confidence interval, 91% to 100% and 92% to 100%, respectively). The procedure was validated in a series of 105 patients with AML. The method confirmed all translocations detected using standard cytogenetics and fluorescence in situ hybridization and some additional undetected rearrangements. Two patients demonstrated two molecular rearrangements simultaneously, with BCR-ABL1 implicated in both, in addition to RUNX1-MECOM in one patient and PML-RARA in another. In conclusion, this novel real-time RT-PCR assay for simultaneous detection of multiple AML-associated fusion genes is a versatile and sensitive method for reliable screening of recurrent rearrangements in AML.
authors.affiliation: Laboratory of Molecular Biology, Department of Clinical Chemistry, University Hospital La Fe, Valencia, Spain.
authors.forename: Sandra
authors.initials: S
authors.lastname: Dolz
authors.forename: Eva
authors.initials: E
authors.lastname: Barragán
authors.forename: Óscar
authors.initials: Ó
authors.lastname: Fuster
authors.forename: Marta
authors.initials: M
authors.lastname: Llop
authors.forename: José
authors.initials: J
authors.lastname: Cervera
authors.forename: Esperanza
authors.initials: E
authors.lastname: Such
authors.forename: Inmaculada
authors.initials: I
authors.lastname: De Juan
authors.forename: Sarai
authors.initials: S
authors.lastname: Palanca
authors.forename: Rosa
authors.initials: R
authors.lastname: Murria
authors.forename: Pascual
authors.initials: P
authors.lastname: Bolufer
authors.forename: Irene
authors.initials: I
authors.lastname: Luna
authors.forename: Inés
authors.initials: I
authors.lastname: Gómez
authors.forename: María
authors.initials: M
authors.lastname: López
authors.forename: Mariam
authors.initials: M
authors.lastname: Ibáñez
authors.forename: Miguel A
authors.initials: MA
authors.lastname: Sanz
mesh_terms: D000293:Adolescent; D000328:Adult; D000368:Aged; D002648:Child; D020732:Cytogenetic Analysis / Q000379:methods; D005260:Female; D006801:Humans; D015470:Leukemia, Myeloid, Acute / Q000175:diagnosis* / Q000235:genetics*; D008297:Male; D008875:Middle Aged; D015514:Oncogene Proteins, Fusion / Q000235:genetics*; D060888:Real-Time Polymerase Chain Reaction / Q000379:methods / Q000592:standards; D015203:Reproducibility of Results; D012680:Sensitivity and Specificity; D014178:Translocation, Genetic; D055815:Young Adult
chemical_list: D015514:Oncogene Proteins, Fusion